1. Field of Industrial Application
This invention relates to an immunological assay method. More particularly, it relates to an immunological assay method suitable for quantitatively determining an antigen or an antibody.
2. Prior Art
Known methods for assaying bodily fluid components such as IgG, IgA, Igm, complements C3 and C4 and CRP include nephelometry with the use of an autoanalyzer provided with a built-in computer, latex nephelometry and laser light scattering nephelometry.
Each of these methods comprises reading a change in optical properties caused by an antigen/antibody reaction in a solution in terms of a change in transmitted or scattered light and comparing the obtained data with a calibration curve, which has been formed each time by assaying a standard material of a known concentration, to thereby determine the concentration of the substance to be assayed.
Now operations of the conventional nephelometry with the use of an autoanalyzer will be described in detail.
1. A reagent to be assayed, i.e., a sample is introduced into a container such as a cuvette and incubated in an autoanalyzer at an appropriate temperature, for example, 37.degree. C. PA1 2. A calibration curve is formed with the use of a standard material. PA1 2-1. A specific equation, for example, Y=Ax+B/x inputted in a built-in ROM or microcomputer in the analyzer according to the item to be assayed is called. PA1 2-2. The parameters A and B in the above equation are determined depending on the assay of the standard material. PA1 2-3. A calibration curve is formed by substituting the parameters A and B as defined above in the above equation. PA1 3. The sample is measured and the concentration of the antigen or antibody therein is determined according to the above calibration curve. PA1 1. Preparing a reagent. PA1 2. Forming a calibration curve with the use of, for example, a graph and determining an equation corresponding to the calibration curve, for example, Y=Ax+B/x, which should be informed to a user. PA1 3. Prior to the assay of a sample, assaying a standard material of a known concentration. Based on the results thus obtained, determining the parameters in the above equation and forming a calibration curve therefrom. PA1 4. Effecting a series of continuous assays depending on the calibration curve as formed above. PA1 5. Repeating the procedures of steps 3 and 4 each time assays are newly initiated. PA1 1. A sample to be assayed is introduced into a container such as a cuvette and incubated in an autoanalyzer at an appropriate temperature, for example, 37.degree. C. PA1 2. A calibration curve is formed. PA1 2-1. A specific equation, for example, Y=Ax+B/x inputted in a built-in ROM or microcomputer in the analyzer according to the item to be assayed is called. PA1 2-2. A calibration curve is formed by inputting parameters, e.g., A and B, which are recorded in a magnetic medium attached to the reagent kit. PA1 3. A sample is assayed to thereby determine the concentration of an antigen or an antibody. PA1 1. Preparing a reagent. PA1 2. Forming a calibration curve with the use of a standard material of a known concentration. Namely, forming an equation corresponding to the item to be assayed, for example, Y=Ax+B/x, and inputting the same to a computer. Alternately determining parameters corresponding to the reagent, for example, A and B. PA1 b 3. Recording the information concerning the calibration curve thus formed or the parameters thus determined in a recording medium selected from among: PA1 1. When no calibration curve has been formed by the manufacturer in the preparation of the reagent, inputting the preliminarily determined parameters to thereby give a calibration curve corresponding to the reagent. PA1 2. Discontinuously assaying samples to thereby determine the concentrations of the substance to be assayed according to the calibration curve as formed above, without forming any additional one prior to each assay.
When a number of samples are to be assayed, 400 to 500 samples are continuously assayed in a day in general. In the conventional method as described above, it is required to form a new calibration curve each time prior to the initiation of the assay when a series of continuous assays have been completed and the operations are to be carried out again on the next day to approximately one week thereafter.
Thus these operations should be shared between a reagent manufacturer and a user in the following manner.
Reagent manufacturer:
User:
In these conventional methods, which are called calibration systems, it is troublesome to form a calibration curve each time when a few samples are to be assayed or samples should be emergently assayed. Further it is difficult to efficiently utilize a reagent in these cases, since a calibration curve should be formed prior to each assay with the use of a standard material.